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Topic:Reproduction
Reproduction in horses encompasses the biological processes and mechanisms involved in the breeding and development of equine offspring. This includes the study of reproductive anatomy, physiology, and endocrinology in both mares and stallions. Key areas of interest include the estrous cycle, ovulation, conception, gestation, and parturition. Researchers also examine factors influencing fertility, reproductive technologies such as artificial insemination and embryo transfer, and management practices that impact reproductive success. This page compiles peer-reviewed research studies and scholarly articles that explore the physiological, genetic, and environmental aspects of equine reproduction.
Motility, morphology and triple stain analysis of fresh, cooled and frozen-thawed stallion spermatozoa. The aim of the present study was to determine whether there are characteristics of fresh, cooled and frozen-thawed semen samples that can be used to predict the suitability of stallion semen for preservation by cooling or freezing. Each of three ejaculates obtained from 12 stallions was divided into aliquots to be analysed for sperm motility, morphology and membrane integrity as fresh, cooled and frozen-thawed samples. The percentage of morphologically normal spermatozoa was similar in fresh and cooled samples and both were greater than in the frozen samples. There were no strong linear relati...
The role of endogenous opioids in the ovulatory LH surge in mares. Removal of opioid inhibition of GnRH neurones is thought to be a critical event in generating the ovulatory surge in some species. In the present study, a nonsurgical technique was used to collect pituitary venous blood samples from eight mares every 0.5-1.0 min for 1 h before and after administration of the opioid receptor antagonist naloxone (0.2 mg kg(-1), i.v.), to investigate whether opioid inhibition is also important in mares. Jugular blood samples were taken at 10-15 min intervals. Mares were studied 0, 1 or 2 days before ovulation. Naloxone administration increased mean rates of GnRH ...
Partial characterization of an equine conceptus prostaglandin inhibitory factor. Equine conceptuses are thought to produce antiluteolytic factors that inhibit endometrial PGF2alpha and, thus, prevent luteolysis in pregnant mares. The aim of the present study was to characterize partially the chemical nature of the prostaglandin inhibitory factor (PIF) produced by equine conceptuses in vitro. Embryos were collected from pregnant mares 13 +/- 0.5 days after ovulation and were cultured for 24 h. Harvested equine conceptus conditioned media (CCM) were assayed for antiluteolytic activity by determining the inhibition of endometrial PGF2alpha synthesis in vitro. Significant anti...
Relationship between thyroid function and seasonal reproductive activity in mares. The relationship between thyroid function and seasonal reproductive activity in mares was investigated by comparing plasma triiodothyronine and thyroxine concentrations in anoestrous mares and in mares with cyclic ovarian activity during the anovulatory season. In study 1, the diurnal changes in plasma triiodothyronine and thyroxine concentrations on a single day were compared in anoestrous and cyclic mares. The mean thyroxine concentrations were significantly higher (P < 0.001) in the cyclic mares than in the anoestrous mares, whereas mean triiodothyronine concentrations were similar in bo...
Apoptosis in equine granulosa cells and its relationship to cumulus expansion and oocyte chromatin configuration in ovarian follicles. During the oestrous cycle follicles grow and either ovulate or regress. Regressing follicles undergo atresia and in many species apoptosis has been identified as the underlying mechanism in this process. The aims of this study were to establish whether equine granulosa cells degenerate via an apoptotic mechanism and whether the presence of apoptotic cell death in granulosa cells is correlated with oocyte quality. Ovaries from mares at unknown stages of the oestrous cycle were obtained from an abattoir. In Expt 1, follicles (n=352) from 37 mares were processed. DNA was extracted from granulosa ...
Ovulation synchrony after follicle ablation in mares. Two experiments were performed to determine the efficacy of ultrasound-guided transvaginal follicle ablation for synchronizing ovarian function in mares. The experiments were initiated at random stages of the oestrous cycle in control (nonablated) and follicle-ablated mares. On day 0, all follicles > or =10 mm in diameter were punctured, aspirated and curettaged in ablated mares, and, on day 4, two doses of PGF2alpha were administered with a 12 h interval between the doses to both ablated and nonablated (control) mares. In Expt 1, hCG was administered to the ablated mares on the first or se...
Indirect determination of stallion sperm capacitation based on esterase release from spermatozoa challenged with lysophosphatidylcholine. A spectrophotometric assay was developed to measure the amount of esterase released from stallion spermatozoa. This assay was used to determine the percentages of capacitated stallion spermatozoa, determined by the ability of spermatozoa to undergo an acrosome reaction and release esterase in response to a lysophosphatidylcholine challenge, for spermatozoa incubated under conditions to increase intracellular calcium and cAMP. Incubation with 100 nmol calcium ionophore A23187 l(-1) induced 66% of stallion spermatozoa to capacitate after 60 min of incubation at 37 degrees C. Subsequent experimen...
Functional disturbances in the endometrium of barren mares: a histological and immunohistological study. Different types of endometrial maldifferentiation were investigated by histopathological (haematoxylin-eosin (HE), periodic acid Schiff-alcian blue and picro-sirius red staining) and immunohistological (oestrogen and progesterone receptors, Ki-67 antigen, intermediate filaments and laminin) analysis of endometrial biopsy specimens from mares (n=34), most of which had been barren for at least 1 year. Two major features were observed: (i) unequal differentiation, in which two functional stages are visible within a single biopsy sample: areas which are differentiated in accordance with the stage ...
Oxytocin and PGF2alpha release in mares resistant and susceptible to persistent mating-induced endometritis. Mares resistant (n=7) and susceptible (n=9) to persistent mating-induced endometritis were artificially inseminated (AI) during oestrus with chilled extended semen. Blood samples were collected from 30 min before AI, for 2 h after AI and again between 16 and 18 h after AI. Samples were assayed for oxytocin and the PGF2alpha metabolite 13,14-dihydro-15-keto PGF2alpha (PGFM). AI caused a significant increase in oxytocin concentrations in both the resistant and susceptible mares and there was no significant difference in oxytocin release between the two groups of mares. Mean PGFM concentrations w...
Effects of steroids on endometrial oxytocin mRNA production. In this study, the roles of oestrogen and progesterone in the regulation of oxytocin gene expression in equine endometrium were examined. Anoestrous mares (n=19) were assigned randomly to one of the following treatment groups: control (vehicle control for 1 day; n=3); progesterone (250 mg progesterone per day for 6 days; n=4); oestradiol (5 mg beta-oestradiol 17-valerate per day for 6 days; n=4); oestradiol plus short duration progesterone (5 mg beta-oestradiol 17-valerate per day for 6 days followed by 250 mg progesterone per day for 6 days; n=4); and oestradiol plus long duration progesteron...
Effects of follicular fluid or progesterone on in vitro maturation of equine oocytes before intracytoplasmic sperm injection with non-sorted and sex-sorted spermatozoa. In Expt 1, compact cumulus oocyte complexes (COCs) were matured in: (i) control medium (Hepes-buffered TCM-199 with 10% oestrous cow serum (OCS) + oestradiol, LH and FSH); (ii) Hepes-buffered TCM-199 with 20% follicular fluid; or (iii) control medium containing 250 ng progesterone ml(-1). Mature oocytes were collected by transvaginal aspiration as a positive control for the in vitro maturation (IVM) treatments. Oocytes were fertilized by ICSI and cultured in Menezo's B2 + 5% fetal calf serum (FCS). There were no significant differences among IVM treatments. In Expt 2, oocytes with expanded COC...
Estimation of sodium and potassium concentrations in the uterine fluid of mares by microdialysis and ion chromatography. Equine blastocyst fluid has a lower [Na+], a higher [K+] and a lower osmolality than does normal blood serum. Based on the assumptions that the sodium pump is primarily responsible for fluid accumulation and that ions transported actively into the blastocyst increase blastocyst osmolality above that of the external medium, we hypothesized that the [Na+] and the osmolality of mare uterine fluid are lower than those of blastocyst fluid. Microdialysis and ion chromatography were used to estimate [Na+] and [K+] of uterine fluid. Mares (n=10) were used for in vivo measurements at different stages o...
The uterus as a source of oxytocin in cyclic mares. In a recent study, continuous administration of oxytocin by subcutaneous minipump to mares from day 8 to 20 after ovulation prevented luteolysis in most of the treated but none of the control mares, indicating a role for oxytocin in cyclical luteolysis in mares. In the present study, measurement of oxytocin concentrations in uterine flushings recovered from nine mares during days 14-18 after ovulation gave values that were many times higher than those measured concurrently in peripheral plasma. Furthermore, intrauterine administration of oxytocin to four mares on day 14 after ovulation stimula...
The structural morphology and epithelial association of spermatozoa at the uterotubal junction: a descriptive study of equine spermatozoa in situ using scanning electron microscopy. Uterotubal junction tissues (n=12) recovered from young nulliparous mares 4 h after insemination were examined for spermatozoa in situ using scanning electron microscopy. Mares were inseminated before ovulation with semen containing 66-85% morphologically abnormal spermatozoa and approximately 1 x 10(9) progressively motile spermatozoa. Spermatozoa were found in all the tissues and displayed a variety of relationships to the epithelium, depending on regional location, spatial constraints and epithelial surface characteristics. Spermatozoa were most abundant in the epithelial folds on the uteri...
Simultaneous recording of pituitary oxytocin secretion and myometrial activity in oestrous mares exposed to various breeding stimuli. The aim of this study was to investigate the effect of different breeding stimuli on uterine contractility and pituitary oxytocin release in five oestrous mares, in order to design better treatments for mares with defective physical uterine clearance mechanisms. Electrodes and strain gauges were implanted surgically on the uterine myometrium and myometrial activity was monitored with a Grass polygraph. A catheter was placed non-surgically in the intercavernous sinus of each oestrous mare to sample pituitary venous blood and a second catheter was placed in the jugular vein. Continuous sampling ...
The role of seminal plasma in post-breeding uterine inflammation. The effect of seminal plasma on polymorphonuclear neutrophil (PMN) chemotaxis, PMN phagocytosis and complement-induced cytolysis was determined using blood plasma pooled from four horses and seminal plasma pooled from two stallions. To investigate chemotaxis, complement in blood plasma was activated with E. coli lipopolysaccharide in the presence of 0-50% seminal plasma diluted with a standardized volume of McCoy's medium and placed in a chemotactic chamber. Chemotaxis of blood derived equine PMNs toward the chemoattractants was determined after incubation at 37 degrees C for 45 min. To invest...
Preservation of stallion sperm quality by native phosphocaseinate: a direct or indirect effect? Milk-based diluents are generally considered efficient for survival of stallion spermatozoa in vitro. However, milk is a complex and variable medium and native phosphocaseinate is a milk component that is more efficient for preservation of sperm motility and fertility, although the mechanisms involved in this protection have not yet been elucidated. The aim of the present study was to characterize the interactions between native phosphocaseinate and equine spermatozoa. No binding between sperm membranes and native phosphocaseinate was observed using indirect immunofluorescent staining or elect...
Effect of increased daylight during late pregnancy on the reproductive performance of mares after parturition. The aim of the present study was to determine the effect of prolonged photoperiod during late pregnancy on subsequent ovarian activity and fertility in mares. Pregnant mares (n=13) due to give birth in January and February were stimulated by a fixed photoperiod (16 h light: 8 h dark) from 15 November (during the last 2-3 months of gestation) until up to 1 month after parturition. A control group of mares (n=9) due to give birth at the same time were kept in the same stable and management regimen, but under natural light conditions. Light-treated mares ovulated during foal oestrus approximately...
Possible role of autoimmunity to spermatozoa in idiopathic infertility of stallions. Testicular degeneration is a major cause of subfertility in stallions, although an aetiological diagnosis cannot be made in most cases. In the present study, autoimmune testicular degeneration was induced and evaluated in stallions by immunizing stallions with their own spermatozoa mixed with an adjuvant. The factors evaluated included changes in semen quality and testicular histology. A large decrease in sperm number and quality was observed in response to sperm autoantigens. An ELISA test specific for antisperm antibodies was developed which enabled antibody titres in serum, seminal plasma a...
Mucus production by endometrium of reproductively healthy mares and mares with delayed uterine clearance. The aims of this study were to compare various staining and fixation techniques for endometrial biopsy samples and, thus, to investigate the mucociliary apparatus in endometrium from reproductively healthy mares and mares with delayed uterine clearance. Endometrial samples were collected from the left and right uterine horns of reproductively healthy mares (n=5) and mares with delayed uterine clearance (n=4) during anoestrus, transition, oestrus and dioestrus. Each sample of endometrium was fixed in either Bouin's fixative or formalin, and stained with alcian blue (pH 1.0 or 2.5), periodic aci...
Production of live foals from sperm-injected oocytes harvested from pregnant mares. In vitro fertilization in horses has been less successful than anticipated owing to: (i) the inability to collect large numbers of good quality oocytes; (ii) alterations in the zona pellucida that occur during in vitro maturation of equine oocytes; and (iii) inadequate preparation of equine sperm cells. In addition, studies in humans, mice and cattle have indicated that high concentrations of glucose in culture media may inhibit embryonic development in vitro and this may also be a problem for development of equine embryos in vitro. The aims of the present study were: (i) to achieve fertilizat...
Relationship between the degree of endometrial periglandular fibrosis and the presence of angiotensin-converting enzyme in the equine endometrium. Endometrial periglandular fibrosis (EPF) has been proposed as a possible aetiology for equine embryonic and fetal loss. However, the pathophysiology of EPF is not well understood. Angiotensin-converting enzyme (ACE) is found in macrophages, endothelium (during angiogenesis) and myofibroblasts at sites of fibrosis in the heart, kidneys, liver and skin in several species. An increase in local tissue ACE-binding activity appears to be a critical event in the initiation and progression of fibrosis in these tissues. The aim of this study was to investigate the correlation between ACE activity in th...
Comparison of the cryoprotectant properties of glycerol and ethylene glycol for early (day 6) equine embryos. Early (day 6) equine embryos (n=23) were assigned to four treatment groups to assess the cryoprotectant properties of glycerol and ethylene glycol and the effect of adding sucrose during removal of the cryoprotectant: (i) group GG (n=5) embryos were frozen and thawed using 1.5 mol glycerol l(-1) as the cryoprotectant, which was added at 22 degrees C in four steps (0.375, 0.75, 1.125 and 1.5 mol glycerol l(-1)), and removed after thawing in five steps (1.5, 1.125, 0.75, 0.375 and 0.0 mol glycerol l(-1)); (ii) group GS (n=6) embryos were frozen and thawed using 1.5 mol glycerol l(-1) as for grou...
Simultaneous exocrine and endocrine secretion: trophoblast and glands of the endometrial cups. Invasion of trophoblast cells of the chorionic girdle into the endometrium and their subsequent dependence as endometrial cup cells on the maternal vascular system are unique features of equine gestation. This study of exocrine secretion by the endometrial glands and endocrine secretion by the cup cells was initiated to examine the relationships between and the effects of two disparate epithelial cell types sharing the endometrium. Endometrial cups were examined from day 36 to day 83 of gestation by light and electron microscopy, and immunohistochemical and lectin analysis. As the cup cells in...
A detailed study of hormonal profiles in mares at luteolysis. Jugular blood samples were collected at 4 h intervals from six mares during an oestrous cycle to study the hormonal events that occur around the time of luteolysis. Blood samples from day 10 (day 0 = ovulation) until day 3 of oestrus were assayed for prostaglandin metabolite 13,14-dihydro-15-keto PGF2alpha (PGFM), oxytocin, prolactin, progesterone and oestrogen conjugates. PGF2alpha (0.5 or 1.5 mg) was administered to six mid-dioestrous mares and the oxytocin and prolactin responses were measured. One to five large (peak > or =2 x nadir) pulses of PGFM, oxytocin and prolactin were detected ...
Progesterone, oestrogen and glucocorticoid receptors in the uterus and mammary glands of mares from mid- to late gestation. Progesterone, oestrogen and glucocorticoid receptor concentrations in the uterus and mammary glands of pregnant mares during mid- to late gestation (from day 150 of gestation to term) were measured by binding assays to investigate the hormonal mechanisms involved in pregnancy maintenance and lactation. Uterine progesterone receptor concentrations did not increase significantly with increasing gestational age (from 67.8 +/- 13.7 to 126.1 +/- 48.7fmol mg(-1) protein), whereas oestrogen receptor concentrations were significantly higher in pregnant mares (271.7 +/- 28.9 fmol mg(-1) protein) than i...
Transabdominal ultrasonographic determination of fetal gender in the horse during mid-gestation. Gender determination of the equine fetus using transabdominal ultrasonography was studied in 20 mares. One group of 10 research mares was scanned repeatedly every 2 weeks from 100 days gestation to parturition, while the second group of 10 client mares was subjected to echography once during mid-gestation. In males, the penis and/or prepuce was observed on 71 occasions from 102 days to 258 days gestation. On cross-sectional views, the male external genitalia had a round shape with parallel linear echogenic foci up to approximately 140 days gestation and then appeared triangular. Fetal testes w...
Measurements of reproductive function in stallions treated with trimethoprim-sulfamethoxazole and pyrimethamine. To evaluate the effects of trimethoprim-sulfamethoxazole and pyrimethamine treatment on various measures of reproductive function in healthy pony stallions. Methods: Randomized complete block study. Methods: 12 healthy, mature pony stallions. Methods: Stallions were assigned to treatment and control groups balanced for age and various characteristics of reproductive function. The treated group received trimethoprim-sulfamethoxazole and pyrimethamine for 90 days during summer and fall; the control group was not treated. Semen characteristics, sexual behavior, testicular volume, and sperm produc...
Secular sperm trends in stallions between 1981 and 1996. Several reports have suggested that human semen quality has declined throughout the world over the last few decades. Chemicals in the environment acting as endocrine disrupters have been implicated as a possible cause. If this is indeed the case, then similar effects may be observed in animals. We analyzed 1489 ejaculates collected from 390 Breton draught stallions between 1981 and 1996. Semen was collected from all the stallions at a single center, according to standardized semen collection protocols and laboratory methods. Semen volume decreased slightly but significantly and there was an in...
Equine reproductive pharmacology. This article reviews therapies and strategies commonly used to treat diseases of the mare's genital tract and modulate the reproductive cycle of the mare. Many reproductive treatments are based on historical use and empirical evidence rather than well controlled clinical studies. This article attempts to present practical information in a summary form while highlighting the need for continued research documenting the efficacy and safety of reproductive therapies.
